His5

Abstract

The invention provides HIS5 polypeptides and DNA (RNA) encoding HIS5 polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing HIS5 polypeptides to screen for antibacterial compounds.

What is claimed is:  
     
       1. An isolated polynucleotide segment, comprising a first polynucleotide sequence, or the full complement of the entire length of the first polynucleotide sequence, wherein the first polynucleotide sequence encodes the same polypeptide, expressed by the HIS5 gene encoded by a nucleic acid sequence comprising SEQ ID NO:1 contained in  Staphylococcus aureus  WCUH 29 contained in NCIMB Deposit No. 40771. 
     
     
       2. The isolated polynucleotide segment of claim  1  encoding fusion polypeptide wherein the first polynucleotide sequence encodes part of the fusion polypeptide. 
     
     
       3. An isolated polynucleotide segment, comprising a first polynucleotide sequence or the full complement of the entire length of the first polynucleotide sequence, wherein the first polynucleotide sequence hybridizes to the full complement of SEQ ID NO:1, wherein the hybridization conditions include incubation at 42° C. in a solution comprising: 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 micrograms/ml denatured, sheared salmon sperm DNA, followed by washing in 0.1×SSC at about 65° C.; and wherein the first polynucleotide sequence is identical to SEQ ID NO:1 except that, over the entire length corresponding to SEQ ID NO:1, there are up to five point mutations for every 100 nucleotides of SEQ ID NO:1, wherein each point mutation is the substitution, deletion or insertion of one nucleotide; and wherein the first polynucleotide sequence detects  Staphylococcus aureus.    
     
     
       4. The isolated polynucleotide segment of claim  3 , wherein the first polynucleotide sequence hybridizes to the full complement of SEQ ID NO:1, wherein the hybridization conditions include incubation at 42° C. in a solution comprising: 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 micrograms/ml denatured, sheared salmon sperm DNA, followed by washing in 0.1×SSC at about 65° C.; and wherein the first polynucleotide sequence is identical to SEQ ID NO:1 except that, over the entire length corresponding to SEQ ID NO:1, there are up to three point mutations for every 100 nucleotides of SEQ ID NO:1, wherein each point mutation is the substitution, deletion or insertion of one nucleotide. 
     
     
       5. An isolated polynucleotide segment comprising, a first polynucleotide sequence, wherein the first polynucleotide sequence encodes a polypeptide comprising the amino acid sequence as set forth in SEQ ID NO:2. 
     
     
       6. A vector comprising the isolated polynucleotide segment of claim  5 . 
     
     
       7. An isolated host cell comprising the vector of claim  6 . 
     
     
       8. A process for producing a polypeptide, comprising culturing the host cell of claim  7  under conditions sufficient for the production of the polypeptide. 
     
     
       9. The isolated polynucleotide segment of claim  5 , wherein the first polynucleotide sequence encodes a polypeptide consisting of the amino acid sequence as set forth in SEQ ID NO:2. 
     
     
       10. A vector comprising the isolated polynucleotide segment of claim  9 . 
     
     
       11. An isolated host cell comprising the vector of claim  10 . 
     
     
       12. A process for producing a polypeptide, comprising culturing the host cell of claim  11  under conditions sufficient for the production of the polypeptide. 
     
     
       13. An isolated polynucleotide of the formula: 
       
         
           X—(R 1 ) n —(SEQ ID NO:1)—(R 2 ) n —Y  
         
       
       wherein, at the 5′ end of the molecule, X is hydrogen, and at the 3′ end of the molecule, Y is hydrogen or a metal, R 1  and R 2  is any nucleic acid residue, and n is an integer between 1 and 1000. 
     
     
       14. An isolated polynucleotide segment comprising a first polynucleotide sequence or the full complement of the entire length of the first polynucleotide sequence wherein the first polynucleotide sequence is selected from the group consisting of 
       (a) a polynucleotide comprising the nucleotide sequence as set forth in SEQ ID NO:1; and  
       (b) a nucleic acid sequence identical to the polynucleotide of (a) except that, over the entire length corresponding to the polynucleotide of (a), there are up to five point mutations for every 100 nucleotides of the polynucleotide of (a), wherein each point mutation is the substitution, deletion or insertion of one nucleotide  
       ; and wherein the first polynucleotide sequence detects  Staphylococcus aureus.    
     
     
       15. A vector comprising the isolated polynucleotide segment of claim  14 . 
     
     
       16. An isolated host cell comprising the vector of claim  15 . 
     
     
       17. The isolated polynucleotide segment of claim  16 , wherein the first polynucleotide sequence is a nucleic acid sequence identical to the polynucleotide of (a) except that, over the entire length corresponding to the polynucleotide of (a), wherein each point mutation is the substitution, deletion or insertion of one nucleotide. 
     
     
       18. An isolated polynucleotide segment comprising a first polynucleotide sequence or the full complement of the entire length of the first polynucleotide sequence, wherein the first polynucleotide sequence comprises SEQ ID NO:1. 
     
     
       19. A vector comprising the isolated polynucleotide segment of claim  18 . 
     
     
       20. An isolated host cell comprising the vector of claim  19 . 
     
     
       21. A process for producing a polypeptide comprising culturing the host cell of claim  20  under conditions sufficient for the production of the polypeptide, wherein the isolated polynucleotide segment comprises the first polynucleotide sequence. 
     
     
       22. The isolated polynucleotide segment of claim  18  encoding a fusion polypeptide wherein the first polynucleotide sequence encodes part of the fusion polypeptide.